Department of Pharmacology, Faculty of Medicine, Khon Kaen University, Mitraparb Road, Khon Kaen 40002, Thailand.
BMC Cancer. 2013 Dec 5;13:571. doi: 10.1186/1471-2407-13-571.
Phenethyl isothiocyanate (PEITC) is a cancer chemopreventive agent from cruciferous vegetables. Cholangiocarcinoma (CCA) is a chemo-resistant cancer with very poor prognosis. We evaluated the effects of PEITC on induction of apoptotic cell death in relation to cellular glutathione (GSH) and mitochondrial function of a CCA cell line, KKU-M214.
Cytotoxic effects of PEITC on a CCA cell line, KKU-M214, and a reference cell line, Chang cells were evaluated. To delineate mechanisms of cell death, the following parameters were measured; GSH and superoxide levels as the oxidative status parameters, apoptosis related proteins levels using Western blotting. Cellular free calcium level and mitochondrial transmembrane potential were also measured.
PEITC induced apoptotic cell death of both KKU-M214 and Chang cells. After PEITC treatment, both cells showed decrease of Bcl-xl and increase of Bax levels. While KKU-M214 cells released AIF, Chang cells released cytochrome c, with subsequent activation of caspase 3 and 9, upon PEITC treatment. PEITC induced superoxide formation in both cells, although it seemed not play a role in cell death. PEITC caused GSH redox stress in different ways in two cell types, because N-acetylcysteine (NAC) prevented redox stress in Chang but not in KKU-M214 cells. The loss of mitochondrial transmembrane potential was induced by PEITC concurrent with GSH stress, but was not a primary cause of cell death. The rapid increase of free calcium level in cytosol was associated with cell death in both cell lines. These events were prevented by NAC in Chang cells, but not in KKU-M214 cells.
PEITC induced cell death KKU-M214 cells and Chang cells via increase of cellular calcium mobilization and activation of mitochondrial cell death pathway. The effects of PEITC on the redox stress was mediated via different ways in CCA and Chang cells because NAC could prevent redox stress in Chang cells, but not in KKU-M214 cells. The multiple effects of PEITC may be useful for the development of novel chemotherapy for CCA.
苯乙基异硫氰酸酯（PEITC）是十字花科蔬菜中的一种癌症化学预防剂。胆管癌（CCA）是一种化疗耐药的癌症，预后非常差。我们评估了 PEITC 对诱导 CCA 细胞系 KKU-M214 细胞凋亡死亡的影响与细胞谷胱甘肽（GSH）和线粒体功能的关系。
评估 PEITC 对 CCA 细胞系 KKU-M214 和参考细胞系 Chang 细胞的细胞毒性作用。为了描绘细胞死亡的机制，测量了以下参数；氧化状态参数的 GSH 和超氧阴离子水平，使用 Western 印迹法测量凋亡相关蛋白水平。还测量了细胞内游离钙水平和线粒体跨膜电位。
PEITC 诱导 KKU-M214 和 Chang 细胞的凋亡细胞死亡。PEITC 处理后，两种细胞的 Bcl-xl 水平降低，Bax 水平升高。虽然 KKU-M214 细胞释放 AIF，但 Chang 细胞释放细胞色素 c，随后在 PEITC 处理后激活 caspase 3 和 9。PEITC 诱导两种细胞中超氧阴离子的形成，尽管它似乎在细胞死亡中不起作用。PEITC 以不同的方式在两种细胞类型中引起 GSH 氧化应激，因为 N-乙酰半胱氨酸（NAC）可防止 Chang 细胞中的氧化应激，但不能防止 KKU-M214 细胞中的氧化应激。PEITC 诱导的线粒体跨膜电位丧失与 GSH 应激同时发生，但不是细胞死亡的主要原因。细胞质中游离钙水平的快速增加与两种细胞系的细胞死亡有关。这些事件在 Chang 细胞中被 NAC 阻止，但在 KKU-M214 细胞中未被阻止。
PEITC 通过增加细胞内钙动员和激活线粒体细胞死亡途径诱导 KKU-M214 细胞和 Chang 细胞死亡。PEITC 对氧化应激的影响在 CCA 和 Chang 细胞中通过不同的方式介导，因为 NAC 可以防止 Chang 细胞中的氧化应激，但不能防止 KKU-M214 细胞中的氧化应激。PEITC 的多种作用可能有助于开发用于治疗 CCA 的新型化疗药物。
